59 results about "SOX2" patented technology

The present disclosure provides compositions and methods for targeting SOX2 antigen to, for example, treat or manage cancer. Provided compositions include binding proteins that are capable of binding to a SOX2 antigen:HLA complex. Also provided are polynucleotides and transgene constructs encoding binding proteins, such as a T cell receptor or a chimeric antigen receptor, that specifically bind to a SOX2 antigen. Such polynucleotides and transgene constructs can be introduced into an immune cell, such as a T cell, and used in immunotherapy in a subject having or at risk for a cancer associated with SOX2 expression or activity. The present disclosure also provides immunogenic compositions comprising SOX2 antigens, and related uses.

The present invention discloses an application of highly active human umbilical cord mesenchymal stem cells. HA-MSCs were isolated from human umbilical cord tissue using TeSR-E8 containing 5% CloneR and LN521-Coated culture flasks, and a technical system for HA-MSC isolation, expansion, and cryopreservation was established. Compared with conventionally cultured MSCs, the HA-MSCs prepared by the present invention have higher proliferation activity, smaller size, a higher nuclear-cytoplasmic ratio, and highly express embryonic stem cell-associated marker antigens SOX2, Nanog, and OCT4. They have the potential to differentiate into neural, myocardial, and hepato-intestinal progenitor cells derived from the three germ layers and possess excellent anti-aging activity.

The invention belongs to the technical field of gene engineering, and particularly relates to a pleural mesothelioma animal model construction method and application capable of simulating immunotherapy response, and the method comprises the following steps: culturing human or mouse lung cells into a first organ, carrying out TrypLE resuspension digestion into single cells, carrying out genetic modification, and culturing into a second organ; the genetic modification is one of the following modes: I, knocking out Trp53, Pten, Cdkn2a and Cdkn2b genes, and overexpressing a Sox2 mutant gene and a Myc gene; and II, knocking out the Trp53 gene, the Pten gene, the Cdkn2a gene, the Cdkn2b gene and the Bap1 gene, and carrying out overexpression on the Sox2 mutant gene and the Myc gene. Resuspending and digesting the second organ to prepare a cell suspension; and injecting the mixture between the dirty layer pleura of the mouse, and culturing for 90-120 days to construct the pleural mesothelioma mouse model. Compared with a transplanted tumor animal model, the method has the advantages that the animal model keeps a natural microenvironment for tumor generation and development in an animal body, and the accuracy is higher in the aspect of simulating real clinical patient conditions.

The invention discloses a biomarker for accurate typing of esophageal squamous carcinoma lesion and application of the biomarker, and belongs to the technical field of molecular biology, the biomarker is SOX2 protein and KDM4B protein which are co-localized in nuclei, and the biomarker is applied to accurate typing of esophageal squamous carcinoma lesion with a non-diagnostic purpose. The cell proportion and distribution conditions of the SOX2 protein and the KDM4B protein are simultaneously displayed on the same section by adopting a double-label immunofluorescence method, and tests prove that the intranuclear co-localization of the SOX2 and the KDM4B can be used as an early warning mark and is used for identifying high-risk patients or early stages of precancerous lesions or distinguishing tumor patients, so that the missed diagnosis rate is reduced, and the diagnosis time is shortened. Therefore, a reliable basis is provided for accurate typing of esophageal squamous carcinoma lesions.

The invention relates to the technical field of biological medicine, and relates to a polypeptide in targeted combination with SOX2 protein and application thereof, the polypeptide has an amino acid sequence as shown in SEQ ID NO.1, or has at least 85%, 90%, 95% or more than 99% of sequence identity with the amino acid sequence; the polypeptide inhibits phase separation of SOX2, liquid drops formed by phase separation of SOX2 can be avoided, then the situation that the liquid drops wrap chemotherapeutic drugs, influence the curative effect of the chemotherapeutic drugs and cause tolerance of tumor cells to the chemotherapeutic drugs is avoided, and the polypeptide does not influence the transcription activation function of SOX2 serving as a transcription factor; the polypeptide can inhibit the phase separation of the SOX2, so that the regulation of normal physiological functions, such as high expression in organs such as eyes of a human body, of the SOX2 as a transcription factor is not influenced, and therefore, the polypeptide disclosed by the invention can reverse chemotherapy drug resistance of tumors such as squamous carcinoma by inhibiting the phase separation of the SOX2, can be used for adjuvant therapy of the tumors such as squamous carcinoma, and has no obvious toxic or side effect on each organ of an organism.

The invention relates to the technical field of cell preparation, and particularly discloses a cell drug and a preparation method thereof.The cell drug comprises mesenchymal cells and pharmaceutically acceptable auxiliary materials; the mesenchymal cell is characterized in that the expression of SOX2, DPPA4 and MYCN genes is negative; the auxiliary material comprises a sodium chloride solution containing albumin; the concentration of the sodium chloride solution is 0.1%-2%; the content of the albumin in the sodium chloride solution is 0.1%-10%. The cell medicine is simple in composition and good in auxiliary material compatibility, the motility of active ingredients can be protected, and the safety and reliability of the cell medicine are remarkably improved by optimizing the preparation process.

The present application discloses cell clusters resembling the function and characteristics of endogenous pancreatic islets, and methods for making and using such cell clusters. The present disclosure provides an in vitro cell cluster comprising at least one non-native pancreatic β cell that exhibits an in vitro glucose-stimulated insulin secretion response when exposed to a glucose challenge, wherein the cell cluster is an unsorted cell cluster, and wherein the cell cluster comprises: at least 35% of cells that express NKX6.1 and C-peptide; at least 70% of cells that express chromogranin A; at most about 2% of cells that express SOX2; or at most about 10% of cells that express SOX9. In some cases, when transplanted into a subject, the in vitro cell cluster exhibits an in vivo glucose-stimulated insulin secretion response to a glucose challenge in the subject.

Disclosed is an efficient and non-genetically modified iPSC-induced, industrialized single clone selection platform, and a use. The platform can efficiently perform reprogramming, and only requires the use of a minimal number of reprogramming factors (OCT4, SOX2, E6, E7). During the single clone separation stage of the present invention, SSEA4 / TRA-1-60 is used as a screening marker, and a large number of single cell clones are obtained by means of flow cytometry. The platform described in the present invention has advantages such as high reprogramming efficiency, high safety, easy operation, and large-scale production.

The application discloses a kind of compositions and applications for regenerating retinal ganglion cells by using endogenous microglia.The present application finds that multiple genes (Brn3b, Sox2, Cbln1 and NP1, referred to as BSCN) can be expressed in microglia to transform into retinal ganglion cells by in vivo microglia fate lineage tracking.The regenerated ganglion cells project axons to the distant brain, rebuild visual neural circuit and restore impaired vision in adult glaucoma mice.In addition, the regenerated ganglion cells can stably survive for up to one year, and the same regeneration strategy is used in old glaucoma mice, which confirms the effectiveness of BSCN reprogramming to regenerate RGCs.Therefore, the present application determines that microglia is a new type of reprogramming seed cell, and finds that four key genes are involved in RGCs regeneration to restore vision, which provides a new therapeutic drug and means for the treatment of glaucoma and related diseases.

The present invention relates to a composition for the prevention or treatment of cisplatin-resistant cancer comprising an inhibitor of DUSP23 expression or activity as an active ingredient. It was found that DUSP23 expression is upregulated in cell clusters exhibiting characteristics of cancer stem cells, and that knocking down DUSP23 significantly reduces cell cluster formation as well as decreases the expression of SOX2, a major stem cell marker. Furthermore, DUSP23 expression was increased in lung cancer cells that had developed cisplatin resistance, and it was confirmed that the downregulation of DUSP23 inhibits self-renewal and invasive capabilities during the induction of cancer cell death. In addition, it was confirmed that DUSP23 plays an important role in promoting cancer stem cell characteristics through SOX2 expression. In conclusion, it has been shown that targeting DUSP23 can be a promising therapeutic strategy to overcome anticancer drug resistance in lung cancer, and the composition of the present invention is expected to be usefully utilized as a treatment for lung cancer cells with cisplatin resistance.

The present invention relates to a single domain antibody specifically binding to STAT3 or an antigen-binding fragment thereof, a fusion protein comprising the single domain antibody or the antigen-binding fragment thereof, and a use of the fusion protein.

The invention discloses a culture medium for goat trophoblast organoid, a culture method and application, and belongs to the field of goat trophoblast organoid culture. The method comprises the following steps: separating trophoblast tissues from a healthy goat placenta which is pregnant for 80-90 days, digesting the trophoblast tissues through type II collagenase to obtain primary cells, compounding the primary cells with matrigel, and performing three-dimensional culture in an improved trophoblast organ culture medium with specific components to form mature organs. The organ is of a typical three-dimensional structure, and KRT23 and SOX2 markers are stably expressed. The organoid culture system based on the goat primary trophoblast cells is established for the first time, gene modification is not needed, the in-vivo physiological status is highly simulated, and the culture system is standardized and good in repeatability. The constructed organoid can be used as an ideal in-vitro model for researching placenta development, evaluating drugs and toxins and modeling infectious diseases in the gestation period, and is of great significance in improving the reproductive research level of goats and the production benefits of animal husbandry.

The present invention relates to the technical field of environmental exposure prevention and treatment. Disclosed is a use of a brain organoid model for a study of simulating brain development damage caused by environmental pollutants in an embryonic period. The brain organoid model at least comprises progenitor cells (SOX2+cells), neurons (MAP2+cells), deep-layer neurons (TBR1+cells) and immature neurons (TUJ1+cells). The simulated exposure of a brain organoid to environmental pollutants constructed in the present invention and subsequent mechanism exploration and therapeutic intervention studies are a promising method for studying the impact of environmental pollutants on human health. Compared with a traditional toxicology testing method, the technology has multiple advantages, helping researchers to better understand the impact of environmental related diseases on human health, and facilitating developing new strategies for preventing and treating these diseases.

The present invention relates to a method for directing differentiation of ventral midbrain NSCs into neurons comprising contacting a cell population comprising ventral midbrain NSCs with an inhibitor of MEK signaling and an inhibitor of NOTCH signaling, wherein the ventral midbrain NSCs co-express the markers FOXA2, LMX1A, EN1, OTX2, 5 and SOX2.

The disclosure relates to a cell culture method for deriving a multipotent neuromesodermal progenitor (NMP) cell and / or lineages of specific anterior-posterior positions along the human body axis and / or cell types of matched anterior-posterior position, from a human pluripotent stem cell (PSC), comprising culturing said stem cell under conditions and with reagents modulating the following signalling pathways: (i) activation of FGF signalling for a duration of about 12-120 hours; (ii) activation of WNT signalling for a duration of about 48-120 hours; (iii) inhibition of SMAD signalling, including TGF-beta / activin-Nodal signalling and BMP signalling, for a duration of about 48-120 hours, wherein said progenitor co-expresses SOX2, TBXT, CDX2 and CDX1 at RNA and / or protein level(s).The disclosure further relates to a composition for use in deriving multipotent NMPs and / or daughter lineages of a specific anterior-posterior position in the human body and / or cell types of matched anterior-posterior position from human pluripotent stem cells (PSC), a multipotent NMP, a cervical and a thoracic NMP obtained by culturing human PSCs, a composition for use in generating lumbar level NMPs from the multipotent NMPs, a lumbar level NMP obtained by culturing the multipotent NMP, a composition for use in generating tbx6 expressing presomitic mesoderm cells from the multipotent NMPs, a tbx6 expressing presomitic mesoderm cell obtained by culturing the multipotent NMP, a human cell derived from the multipotent NMP, a cell population, a kit and a cellular composition.

Animal cells, notably adult fibroblasts, are advantageously reprogrammed in direct lineage reprogramming methods using defined factors to produce proliferative and multipotent induced cardiac progenitor cells (iCPC). The iCPC thus produced can be differentiated under suitable differentiation conditions to cardiac lineage cells including cardiomyocytes, smooth muscle cells, and endothelial cells, as evidenced by expression of lineage specific markers. Sets of factors effective in combination to reprogram the fibroblasts can include a set that includes some or all of 5 factors (Mespl, Baf60c, Nkx2.5, Gata4, Tbx5), a set that includes some or all of 11 factors (Mespl, Mesp2, Gata4, Gata6, Baf60c, SRF, Isll, Nkx2.5, Irx4, Tbx5, Tbx20), a set that includes some or all of 18 factors (T, Mespl, Mesp2, Tbx5, Tbx20, Isll, Gata4, Gata6, Irx4, Nkx2.5, Handl, Hand2, Tbx20, Tbx18, Tip60, Baf60c, SRF, Hey2), and a set that includes some or all of 22 factors (T, Mespl, Mesp2, Tbx5, Tbx20, Isll, Gata4, Gata6, Irx4, Nkx2.5, Hand1, Hand2, Tbx20, Tbx18, Tip60, Baf60c, SRF, Hey2, Oct4, Klf4, Sox2, L-myc).

Methods are provided for generating pancreatic endocrine cells from pluripotent stem cells (PSCs) using epigenetic modifiers. A fusion protein comprising catalytically inactivated Cas9 (dCas9) and an embryonic ectoderm development (EED)-binding domain (EBdCas9) is used with guide RNAs (gRNAs) to target regulatory regions of transcriptional regulators including EOMES, PDX1, and NGN3. Sequential targeting of PDX1 and NGN3 promoters with EBdCas9 removes repressive H3K27me3 marks, increases activating H3K27ac modifications, and accelerates differentiation into insulin-producing β-cells. The methods significantly increase β-cell yields (~4-6 fold), reduce non-pancreatic lineage contaminants (CDX2+, SOX2+, enteroendocrine cells), enhance mitochondrial function, and produce glucose-responsive cells with superior functional maturation in vivo. The epigenetic interventions enforce pancreatic developmental fates while activating NGN3-dependent transcriptional programs necessary for endocrine cell differentiation, insulin secretion, and metabolic maturation. Cells and compositions produced by these methods are also provided.

The invention belongs to the technical field of biology, and relates to application of chimeric polypeptide TAT-Ae alone or in combination with an AKT inhibitor in tumor treatment. Specifically, the invention discloses application of a blocking TRIB3 / AKT inhibitor targeting tumor stemness and proliferation in preparation of drugs for preventing or treating breast cancer independently or in combination with an AKT inhibitor. The amino acid sequence table of the inhibitor for blocking TRIB3 / AKT is SEQ ID No.1, and the AKT inhibitor is Oridonin. The inhibitor for blocking TRIB < 3 > / AKT can be specifically combined with TRIB < 3 >, so that the interaction of TRIB < 3 > / AKT is blocked, the protein degradation of a transcription factor FOXO1 is promoted, and finally the expression of a dry factor SOX2 is reduced; the AKT inhibitor inhibits tumor cell proliferation by inhibiting AKT activity, so that the AKT inhibitor is applied to preparation of drugs for treating and preventing breast cancer, lymphoma, lung cancer and colon cancer. The prepared medicine has the advantages of being remarkable in curative effect, small in toxic and side effect and safe to use when being used for treating tumor diseases.

The present invention belongs to the field of molecular biology technology, and specifically relates to a method for identifying and isolating human hair follicle dermal stem cells. In a first aspect of the present invention, a marker for human hair follicle dermal stem cells is provided, the marker comprising one or more of COL11A1, RBP4, MMP11, and SOX2. By directly obtaining human hair follicle samples, combined with spatial transcriptome and single-cell RNA sequencing technology, the gene expression profile of the human hair follicle dermis is comprehensively captured, thereby identifying the characteristic gene expression of hHFDSC. At the same time, the spatial transcriptome is used to locate the structural position information of hHFDSC and analyze the spatial distribution characteristics of hHFDSC. By combining these two technologies, based on the identified characteristic genes, combined with their spatial localization and differentiation ability, hHFDSC is clearly defined, achieving efficient and accurate analysis of the characteristics and functions of hHFDSC.

The invention discloses application of H3K9me2 as a target spot in preparation of a preparation for inducing stem cells to differentiate to gametes in vitro, and relates to the technical field of stem cell breeding in animal husbandry. An H3K9me2 inhibitor BIX01294 is adopted as a treating agent, and in-vivo treatment is performed on a pregnant mouse and in-vitro treatment is performed on gonad tissue of a female fetal mouse. The result shows that the BIX01294 treatment can cause the reduction of the expression level of H3K9me2, so that the expression levels of pluripotent genes Oct4, Sox2 and Dppa3 are increased, the accessibility of whole genome chromatin is obviously changed, and the meiosis start and meiosis process are blocked. The result shows that the H3K9me2 can influence the differentiation of the stem cells to the gametes and is beneficial to the generation of artificial gametes in the stem cell breeding process.

The invention provides a preparation method of induced pluripotent stem cells, and belongs to the technical field of cell biology. According to the preparation method provided by the invention, the problems of low iPS cell preparation efficiency and high risk in the prior art are solved, the skin fibroblasts can be successfully induced into the induced pluripotent stem cells, and a new means is provided for treatment of related diseases. The invention provides a safer and more efficient technical means, and successfully realizes reprogramming of the skin fibroblasts into the iPS. According to the invention, a small molecule compound is used for replacing retrovirus and lentiviral vectors, and the induced pluripotent stem cells have pluripotent characteristics, including expression of OCT4, SOX2 and Nanog pluripotent markers and formation of teratoma of three-germ-layer tissues. According to the invention, the characteristics of unconformity of small molecules and genomes, high controllability and easiness in optimization, standardization and manufacturing are successfully utilized, so that the obtained chemical reprogramming method is expected to develop a therapeutic reprogramming strategy in a highly controllable and easily standardized manner.

The invention belongs to the technical field of tumor animal models, and particularly relates to a construction method and application of a brain glioma PDTX model. The invention formulates a tumor screening method suitable for constructing the brain glioma PDTX model, tumor samples with relatively high GFAP, SOX2, SLC1A3 and OLIG2 gene transcription levels are screened as ideal tumor sources by performing transcriptome sequencing on tumor tissues, and the model is constructed by the novel brain glioma PDTX model construction method provided by the invention. According to the method, the screened brain glioma tissue is transplanted into the immunodeficient mouse, so that the original microenvironment characteristics of the tumor can be effectively reserved, and the success rate of constructing the brain glioma PDTX model is obviously improved.

The present invention relates to a method of guiding the differentiation of ventral midbrain NSCs into neurons comprising contacting a population of cells comprising ventral midbrain NSCs with an inhibitor of MEK signaling and an inhibitor of NOTCH signaling wherein the ventral midbrain NSCs co-express markers FOXA2, LMX1A, EN1, OTX2 and SOX2.

To provide a separation and discrimination method of a nucleus pulposus precursor cell, and application thereof.SOLUTION: Provided is a separation and discrimination method of a nucleus pulposus precursor cell, where a large number of tissue blocks are divided from a nucleus pulposus tissue, and the tissue blocks are left to stand and cultured in a culture dish after performing hydrolysis by enzyme. When cells are formed into the partition blocks on a bottom of the culture dish, a nucleus pulposus precursor cell is screened from the partition blocks by using a specific embryonic stem cell gene (e.g., Nanog, Oct-4, SOX2). Further, the method can screen a nucleus pulposus precursor cell having moving performance and / or mesenchymal stem cell properties by using a specific protein and a gene marker, and further can screen a nucleus pulposus precursor cell having anti-inflammatory performance. Finally, disclosed is application used for producing a pharmaceutical composition treating lower back pain by using the nucleus pulposus precursor cell obtained by the method, and a new therapy strategy to lower back pain is provided.SELECTED DRAWING: Figure 6

The present invention provides methods of transdifferentiating a somatic cell, e.g., a fibroblast, to an induced dopaminergic precursor cell (iDP) in vitro. The methods include ectopically expressing genes of (1) one or both of Brn2 and Brn4, or a variant thereof, (2) Sox2, or a variant thereof, and (3) one or both of Foxa2 and Lmx1a, or a variant thereof, in the somatic cell, and methods for treating a neurodegenerative disease, such as Parkinson's disease, with the iDP.

Provided are a cow milk-derived stem cell, and an isolation and culture method therefor. The method comprises the following steps: (1) taking fresh cow milk, and isolating cells therefrom; (2) coating a culture dish with a matrix gel in advance, discarding the coating solution, then washing the culture dish with a washing solution, and discarding the washing solution; (3) inoculating the cells treated in the step (1) on the coated culture dish, and culturing the cells with a stem cell culture medium; and (4) when the cell confluence reaches 80% or more, digesting the cells with a digestion enzyme, terminating the digestion, isolating the cells, resuspending the cells using a CTFR culture solution, then inoculating the cells on a feeder layer for culture, promoting colony formation, and performing subculture to obtain cow milk-derived stem cells. The cow milk-derived stem cell has OCT4, SOX2, NANOG and SSEA4 markers.

The invention primarily relates to a method for isolating and characterizing nucleus pulposus progenitor cells. The method initially involves cutting nucleus pulposus tissue into multiple tissue blocks and then enzymatically digesting these blocks. After digestion, these tissue blocks are cultured in a culture dish. When cells form clusters at the bottom of the dish, specific embryonic stem cell genes (such as Nanog, Oct-4, SOX2) are used to select nucleus pulposus progenitor cells from the clusters. Furthermore, this method employs specific proteins and genetic markers to select nucleus pulposus progenitor cells with mobility and / or mesenchymal stem cell characteristics, as well as to further select cells with anti-inflammatory abilities. Finally, the invention discloses a use of the nucleus pulposus progenitor cells obtained by the method in the manufacture of a pharmaceutical composition for the treatment of lower back pain, offering a new strategy for the treatment of lower back pain.

The invention provides a serum-free culture method for converting hUC-MSCs into hiPSCs through induction of a small molecule compound, can be used for preparing pluripotent stem cells, and belongs to the field of biomedicine. Comprising thymine analogs, cAMP activators, TGF-beta receptor inhibitors, bone morphogenetic proteins, RA receptor activators, GSK3 inhibitors and basic fibroblast growth factors, and the culture system does not contain serum. And 2, in the process of carrying out micromolecule compound reprogramming on the hUC-MSCs by utilizing the method provided by the invention, frequent digestion, inoculation and tray separation on the hUC-MSCs are not needed, so that compared with the existing culture method, the operation steps are simplified, the cell loss generated in the digestion, inoculation and tray separation process is reduced, and the cell quality is improved. According to the invention, the reprogramming efficiency of the small molecule compound on hUC-MSCs is obviously improved; and 3, by detecting the co-expression (such as OCT4, SOX2 and NANOG) of transcription factors in reprogrammed cells of a small molecule compound reprogrammed pluripotent stem cell line obtained by the technology, it is proved that the hiPSCs obtained by inducing the hUC-MSCs have stable pluripotency capable of continuous passage.

Provided are materials and methods for rejuvenating senescent cells. Senescent cells are transduced with a viral vector to express Oct4 and Sox2 proteins for partial cell reprogramming and / or to suppress a senescence-associated secretory phenotype. The materials and methods provided can be used to treat progeria syndrome, signs and symptoms of premature aging or natural aging or to rejuvenate tissue in subjects experiencing premature aging or having an agerelated disease.

The present invention relates to a single domain antibody specifically binding to SOX2 via E3 ubiquitin ligase or an antigen-binding fragment thereof, a fusion protein for enhancing proteolysis using same, and a use thereof. The fusion protein comprising the anti-SOX2 single domain antibody and the E3 ubiquitin ligase fragment of the present invention specifically binds to SOX2 to degrade SOX2. SOX2 degradation by the fusion protein promotes growth inhibition and apoptosis of cancer cell lines. In addition, when used in combination with an anticancer agent, the fusion protein inhibited the growth of cancer cells that exhibit drug resistance. The fusion protein inhibits tumor growth in a tumor mouse model. Therefore, the fusion protein of the present invention can be advantageously used for preventing or treating cancer.